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Effects of Estrogen on the Transcriptional Activities of Catecholamine Biosynthesizing Enzymes in the Brain and Adrenal Gland of Ovariectomized Rats (난소 절제 흰쥐의 뇌와 부신에서의 Catecholamine Biosynthesizing Enzyme들의 전사에 미치는 Estrogen의 효과)

  • 유경신;이종화;최돈찬;이성호
    • Development and Reproduction
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    • v.6 no.2
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    • pp.117-122
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    • 2002
  • Dopamine(DA), norepinephrine(NE), and epinephrine(E) belong to a class of neurotransmitters known as catecholamine (CA) which are synthesized and secreted by mammalian brain and adrenal medulla. CA regulate several behavior patterns connected with breeding, and regulate GnRH-gonadotropin hormone axis' vitality between hypothalamus-pituitary gland linking with reproduction freeze. The present study examined effects of sex steroid hormone on the transcriptional activities of CA biosynthesis enzymes, tyrosine hydroxylase(TH), dopamine $\beta$ -hydroxylase(DBH), and phenylethaolamine-N-methyl transferase(PNMT). Mature female rats were ovariectomized(OVX) and implanted with 17 $\beta$-estradiol(E$_2$: 500 $\mu\textrm{g}$/ml) or sesame oil. Forty-eight hours after implantation all the animals were sacrificed. Total RNAs were extracted immediately and were applied to semi-quantitative reverse transcription-polymerase chain reaction(RT-PCR). The expression level of TH was appeared by hypothalamus > SNc> adrenal medulla orders in OVX+Oil group, and by SNc > hypothalamus) adrenal medulla orders in OVX+E$_2$ group. Treatment with E$_2$ significantly increased TH expression in SNc and adrenal medulla but in hypothalamus, the reduced TH expression was observed. The expression level of DBH was appeared by adrenal medulla > SNc > hypothalamus orders in OVX+Oil group and in OVX+E$_2$ group. Administration of E$_2$ significantly reduced DBH expression in SNc, and increased in adrenal medulla. Two cDNA products, large(PNMT1) and small(PMNTs) species of 110bp difference, were amplified in SNc and hypothalamus, but only PNMTs was observed in adenal medulla. The PNMTs expression level was in the order of adrenal medulla > hypothalamus > SNc in both OVX+Oil and OVX+E$_2$ group. The PNMTs expression in SNc and adrenal medulla was significantly increased byE$_2$. The present report demonstrated that estrogen effects on transcriptional activities for CA biosynthethic enzymes were tissue specific in adrenal medulla as well as different region of brain. These results suggest that it might be crucial relationship between the type of estrogen receptor and CA enzyme gene expression.

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Identification and Functional Analysis of Pig β-1,4-N-Acetylglucosaminyltransferase A (MGAT4A) (돼지 유래의 β-1,4-N-acetylglucosaminyltransferase A (MGAT4A) 유전자의 동정 및 기능 분석)

  • Kim, Ji-Youn;Hwang, Hwan-Jin;Chung, Hak-Jae;Park, Mi-Ryung;Byun, Sung June;Kim, Kyung-Woon
    • Journal of Life Science
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    • v.26 no.3
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    • pp.275-281
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    • 2016
  • Glycan modification is important in pharmaceutical industry. Especially, sialic acid affects the bioactivity and stability of medicine. Milk of pig has been used as bioreactor to produce various pharmaceutical proteins. Therefore, it is necessary to modify the glycan chain in pig mammary grand. β-1,4-N-Acetylglucosaminyltransferase A (pMGAT4A) is one of the essential enzymes for increase of sialic acid content, but pig MGAT4A is unclear. In this study, the pMGAT4A was identified and characterized. The pMGAT4A has 1638 nucleotides encoding 535 amino acids and type II membrane topology, which is one of the common features in many glycosyltransferases. The gene was strongly expressed in liver and mammary gland, whereas was weakly expressed in small intestine, stomach and bladder. For functional test, HA-tagged MGAT4A was over-expressed in porcine kidney (PK-15) cell line. Forced expression of pMGAT4A gene was identified by qPCR, and we identified that pMGAT4A is located in Golgi complex by co- staining with HA antibody and BODIPY TR ceramide. In addition, we identified the increase of mannose-β-1,4-N-acetylglucosamine structure by ELISA and immunofluorescence using Datura stramonium agglutinin (DSA), which recognizes mannose-β-1,4-Nacetylglucosamine. Through the specific activity analysis, we showed that pMGAT4A modified bi-antennary to tri-antennary. This event affects sialic acid content. Therefore, we thought that over-expression of pMGAT4A will be necessary in pig mammary grand for improved medicine.

A Study on the Efficacy of Dietary Supplementation of Organic Acid Mixture in Broiler Chicks (육계에 대한 복합 유기산제의 첨가급여 효과에 관한 연구)

  • Kim, Dong-Wook;Kim, Ji-Hyuk;Kim, Sung-Kwon;Kang, Geun-Ho;Kang, Hwan-Ku;Lee, Sang-Jin;Kim, Sang-Ho
    • Journal of Animal Science and Technology
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    • v.51 no.3
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    • pp.207-216
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    • 2009
  • This experiment was conducted to investigate the effects of dietary supplementation of organic acid mixture on growth performance, cecal microflora, blood characteristics and immune response in broiler chicks and to prove the possibility of organic acid mixture as an alternative to antibiotics growth promotor. A total of four hundred eighty, 1-day-old male broiler chicks (Ross$\times$Ross 308) were randomly divided into 4 groups with 4 replicates of 30 birds each. The treatments were NC (free antibiotics), PC (basal diet with virginiamycin 10 ppm and salinomycin 60 ppm), 0.3% organic acid, and 0.5% organic acid. The final body weight and body weight gain were significantly higher in organic acid 0.5% than NC (P<0.05). The feed conversion ratio in all treated groups were significantly improved as compared to that of NC (P<0.05). The carcass rate and relative organs weight were not significantly difference among the groups. The relative weight and length of small intestine in PC were significantly decreased than the other groups. The numbers of cecal coliform bacteria and Salmonella in all treated groups were significantly lower than NC (P<0.05). The number of cecal lactic acid bacteria was not different among the groups. No significant differences among the groups were observed in the contents of total cholesterol, triglyceride, blood urea nitrogen (BUN), albumin, aspartate aminotransferase (AST), and alanine aminotransferase (ALT) in blood serum. The contents of total protein and globulin in blood serum of PC and organic acid treated groups were significantly increased as compared to those of NC (P<0.05). Therefore, albumin:globulin ratio of PC and organic acid treated groups was significantly lower than NC (P<0.05). The total white blood cell (WBC), heterophil, lymphocyte, and stress indicator (heterophil:lymphocyte ratio) were not significantly different among the groups. No significant difference was observed on the expression rate of splenic cytokines mRNA in organic acid treated groups compared to the control. Consequently, supplemental organic acid mixture improved the growth performance, and influenced positive effects on the intestinal microflora by inhibiting the growth of harmful bacteria without any adverse effects on relative weights of organs and blood biochemical parameters in broiler chicks.

Recent Advancement in the Differentiation of Tissues and Organs and Regulation of Gene Expression (조직.기관의 분화와 유전자 발현의 조절, 최근의 진보)

  • Harn, Chang-Yawl
    • Korean Journal of Plant Tissue Culture
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    • v.24 no.1
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    • pp.1-35
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    • 1997
  • Fertilized egg, by successive cell divisions, differentiates into different tissues and organs with various structures and functions. Different cells and tissues contain different proteins, products of selective gene expression. Not all the genes in any genomes are equally active, temporal and spatial gene expression being the general rule. Present paper attempts to review the tanscriptional mechanisms or the initiations of transcription from several angles. In some of the organisms the genes in the process of transcription or the genes in the inactive state can be seen under the light microscope. Some bands of Drosophila polytene chromosomes may exhibit a swollen or puff appearance under certain conditions. A puff, unfolded or decondensed form of chromomere, represents sets of intense transcriptional activity or RNA synthesis. The heterochromatic X chromosome whose genes remain inactive in the female mammals can be visualized as a dark staining structure called Barr body, Configuration of chromatin differs between transcribed and nontranscribed chromatin. Modification to the chromatin facilitates RNA synthesis. The movement of large polymerase molecule along the DNA would probably be facilitated if some modifications of the chromatin configuration is effected. Methylation of cytosines in CG sequences is associated with inactive genes. Methylation can play a role in determination of mammalian cells during embryogenesis. Demethylation is necessary for the gene to be expressed during development A histone modification that is also known to be correlated with transcriptional capacity of chromatin is acetylation of the lysine residues of the core histones. Chromatin containing a high level of histone acetylation is very sensitive to DNase 1. For the transcription to occur TBP must first bind to the TATA box. Another TF, TF IIB, then binds to the promoter-TBP complex, facilitating the access of RNA polymerase to the transcription initiation site. As recently as eight years ago researchers assumed that histones were irrelevant to the regulation of gene expression. Histones combine with the DNA to form nucleosome of the chromatin. Histones are vital participant in gene regulation. Histone and basal factors compete for access to TATA box. When DNA is exposed to basal factors before histones are introduced, the basal factors assemble on TATA boxes preventing the access of histones, allowing transcription to occur, for transcription to begin, activator protein at the upstream activation sequence or enhancer must interact with the tail of histone H4 at TATA box and cause the histone role particle to dissociate from the TATA box leading to partial breakup of the histone core particle and allowing the basal factors to bind to the TATA box. New concept of genomic flux in contrast to the old concept of static genome has been developed based on the powerful new molecular techniques. Genomic changes such as repetitive DNAs and transposable elements, it is assumed but not yet proved, may affect some of the developmental patterns that characterize particular cells, tissues, organs, and organisms. In the last decade or so remarkable achievement have been made in the researches of the structures and functions of TFs and the specific target sequences located in promoters or enhancers where these TFs bind. TFs have independent domains that bind DNA and that activate transcription. DNA binding domain of TFs serves to bring the protein into the right location. There are many types of DNA binding domains. Common types of motifs can be found that are responsible for binding to DNA. The motifs are usually quite short and comprise only a small part of the protein structure. Steroid receptors have domains for hormone binding, DNA binding, and activating transcription. The zinc finger motif comprises a DNA binding domain. Leucine zipper consist of a stretch of amino acids with a leucine residue in every seventh position Two proteins form a dimer because they interact by means of leucine zippers on similar α-helical domain. This positions their DNA binding basic domains for interaction with the two halves of a DNA sequence with dyad symmetry of TGACTCA, ACTGAGT.

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Biodistribution and Hepatic Metabolism of Galactosylated $^{111}In-Antibody-Chelator$ Conjugates: Comparison with $^{111}In-Antibody-Chelator$ Conjugates ($^{111}In$-표지 갈락토즈 접합 항체의 체내분포 및 간에서의 대사 : $^{111}In$-표지 항체와의 비교연구)

  • Kwak, Dong-Suk;Jeong, Kyu-Sik;Ha, Jeoung-Hee;Ahn, Byeong-Cheol;Lee, Kyu-Bo;Paik, Chang-H.;Lee, Jae-Tae
    • The Korean Journal of Nuclear Medicine
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    • v.37 no.6
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    • pp.402-417
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    • 2003
  • Purpose: To evaluate the use of monoclonal antibody (MoAb) as a carrier of the receptor-binding ligand the receptor mediated uptake into liver and subsequent metabolism of $^{111}In-labeled$ galactosylated MoAb-chelator conjugates were investigated and compared with those of $^{111}In$ labeled MoAb. Materials and Methods : T101 MoAb, $IgG_2$ against human lymphocytic leukemic cell, conjugated with cyclic DTPA dianhydride (DTPA) or 2-p-isothiocyanatobenzyl-6-methyl-DTPA (1B4M) was galactosylated with 2-imino-2-methoxyethyl-1-thio-${\beta}$-D-galactose and then radiolabeled with $^{111}In$. Biodistribution and metabolism study was peformed with two $^{111}In-conjugates$ in mice and rats. Results: $^{111}In-labeled$ T101 and its galactosylated conjugates were taken to the liver by the time, mostly within 10 min. However DTPA conjugate was retained longer in the liver than the 1B4M conjugate (55% vs 20% of injected dose at 44 hr). During this time, the radiornetabolite of DTPA conjugate was excreted similarly into urine (24%) and feces (17%). The radiometabolite of 1B4M was excreted primarily into feces (68%) rather than urine (8%). Size exclusion HPLC analysis of the bile and supernatant of liver homogenate showed two peaks the first (35%) with the retention time (Rt) identical to IgG and the second (65%) with Rt similar to free $^{111}In$ at 3 hr post-injection for the 1B4M conjugate, indicating that the metabolite is rapidly excreted through the biliary system. in contrast to DTPA conjugate, the small $^{111}In-DTPA-like$ metabolite was the major radioindium component (90%) in the liver homogenate as early as 3 hour post-injection, but the cumulative radioindium activity in feces was only 17% at 44 hour, indicating that the metabolite from DTPA conjugate does not clear readily through the biliary tract. Conclusion: The galactosylation of the MoAb conjugates resulted in higher hepatocyte uptake and enhanced metabolism, compared to those without galactosylation. Metabolism of the MoAb-conjugates is different between compounds radiolabled with different chelators due to different characteristics of radiometabolites generated in the liver.

Modulation of Cellulalr Quinone Reductase Inducibility by Roasting Treatment and Acid Hydrolysis of Perilla (들깨의 볶음처리와 산가수분해에 의한 세포모델계 Quinone Reductase 활성유도능의 변화)

  • Hong, Eun-Young;Kang, Hee-Jung;Kwon, Chong-Suk;Nam, Young-Jung;Suh, Myung-Ja;Kim, Jong-Sang
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.26 no.2
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    • pp.186-192
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    • 1997
  • Increased activities of phase 2 enzymes including quinone reductase(QR) have been reported to be associated with protection of animals from neoplastic, mutagenic, and other toxic effects of many carcinogens. In previous study, we found that methanol extract of roasted and defatted perilla meal induced the activity of quinone reductase, an anticarcinogenic marker enzyme, in murine hepalc1c7 cells. Current study showed that unroasted perilla had a limited QR-inducing activity, suggesting that roasting cause the generation of active component(s). Thus we hypothesized that QR inducer in perilla might be covalently linked to sugar moiety and released during roasting process. Methanol extract of defatted raw perilla was subject to acid treatment in order to hydrolyze the potential sugar moiety. Prolonged hydrolysis of methanol extract of defatted raw perilla at $98{\sim}100^{\circ}C$ increased the ability to induce cytosolic QR activity of hepalclc7 cells. Furthermore roasting at 180 and $200^{\circ}C$ resulted in significant induction of QR activity. The result strongly support the idea that QR inducer(s) is present in bound form in raw perilla and released during roasting. Cellular QR activity was induced proportionately with the increase of concentration of methanol extract of roasted perilla. The induction of QR by defatted perilla was also examined in the cytosols of liver, small intestine, stomach, lung and kidney of male ICR mice. Induction patterns showed specificity with respect to target tissue and roasting of perilla. Unroasted perilla meal (defatted) significantly induced QR in liver and lung, while roasted perilla meal induced QR in liver and stomach. The observation that raw perilla showed similar QR induction patterns to roasted perilla is consistent with our proposal that QR inducer(s) is present in bound form and released by physical and chemical treatments as digestive or microbial enzymes could release the inducers from inactive glycoside forms in gastrointestinal tract of mice. In conclusion, perilla could exert protective effect against chemically induced carcinogenesis by inducing phase 2 enzymes in biological systems regardless of chemical and physical process such as roasting.

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The Characteristics on the Spatio-temporal Distributions of Phytoplankton Communities in Deukryang Bay, Southwestern Korea (득량만 식물플랑크톤 군집의 시.공간적 분포특성.)

  • 윤양호
    • Korean Journal of Environmental Biology
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    • v.17 no.4
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    • pp.481-492
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    • 1999
  • The observations on the spatio-temporal distribution and seasonal fluctuations of phytoplankton community were carried out in Deukryang Bay of the Korean Southwestern Sea from June 1992 to April 1993. A total of 75 species of phytoplankton belonged to 47 genera was identified. In Deukryang Bay seasonal succession in dominant species; P. alata, G. flaccida, S. costatum, L. danicus and N. longissima in summer, St. palmeriana, Ch. curvisetus and B. paxillifera in autunm, S. costatum, Ch. curvisetus, E. zodiacus and Pn. pungens in winter, and As. glacialis, As. kariana, N. pelagica, Th. nitzschioides and S. costatum in spring, were very marked, that is to say, the communities structure of phytoplankton in Deukryang Bay appeared to be various species composition and it was occupied with diatoms all the year round. Phytoplankton standing crops fluctuated with an annual mean of $1.4{\times}10^5 cells/1 between the lowest value of 2.6{\times}10^3 cells/1 in July and the highest value of 1.0{\times}10^6 cells/1$ by S. costatum in January. Densities of the phytoplankton cell number by the samples of Deukryang Bay ranged from $2.6{\times}10^3cells/1 to 1.2{\times}10^5 cells/1 with the mean value of 3.6{\times}10^4cells/1 in summer, from 6.0{\times}10^3cells/1 to 2.6{\times}10^5 cells/1 with mean of 1.5{\times}10^5 cells/1 in autumn, from 1.3{\times}10^4cells/1 to 1.0{\times}10^6 cells/1 with mean 3.5{times}10^5 cells/1 in winter, and from 4.8{\times}10^3cells/1 to 6.0{\times}10^5 cells/1 with mean of 1.6{\times}10^5 cells/1$ in autumn. That is to say, phytoplankton standing crops was large in low temperature seasons, on the other hand small in high temperature seasons. Chlorophyll $\alpha$ concentration fluctuated between 0.l9 $\mu$g/l and 12.3 $\mu$g/l in March. in Deukryang Bay seasonal flucturation in chi-$\alpha$ concentration was not marked. Especially, chl-$\alpha$ concentration in the water around Deukryang Island located in the middle part of Deukryang Bay showed patchy distributions with a very high concentration. And chl-$\alpha$ concentration was high during a year. Therefore, phytoplankton production in Deukryang Bay could be very high year-round.

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Risk factors of Pneumonectomy in Non-Small Cell Lung Cancer (비소세포폐암에시 전폐절제술의 위험 인자)

  • Hwang Eun-Gu;Baek Heejong;Lee Hae-Won;Park Jong-Ho;Zo Jae-Ill
    • Journal of Chest Surgery
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    • v.38 no.9 s.254
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    • pp.616-621
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    • 2005
  • Background: In the resection of lung cancer, pneumonectomy occupied $20 {\~}35\%$ of all resections, and significantly high operative mortality is reported in right pneumonectomy ($10{\~}25\%$). The aim of this study is to identify the characteristics of morbidity, operative mortality and factors affecting operative mortality after pneumonectomy. Material and Method: This study recruited the database which performed pneumonectomy for lung cancer in Korea Cancer Center Hospital from Aug 1987 to Apr 2002. Result: Total of 386 pneumonectomies were peformed in that period. Sidedness were left in 238, right in 148; and the procedures were standard resection in 207, and extended resection in 179. Morbidity occurred in 115 cases ($29.8\%$, 115/386). Mortality occurred in 12 cases ($3.1\%$, 12 in 386). This mortality rate was similar to that of lobectomy ($2.1\%$, 13 in 613) during the same period. Morbidity consisted of 42 hoarseness, 17 (9) pneumonia and ARDS, 8 empyema, 5 (1) broncho-pleural fistula, 5 reoperation for bleeding, 5 (1) arrhythmia, 1 (1) pulmonary edema, and 25 others (The number in the parenthesis is the number of mortality case for that morbidity). Several factors affecting the operative mortality were evaluated. At first, extended procedure ($3.3\%$, 6 in 179) affected the operative mortality similar to the standard procedure ($2.9\%$, 6 in 207)(p=0.812). Second, the rate of operative mortality in an elderly group over 60 years ($5.5\%$, 10 in 182) was significantly higher than the younger group under 60 years ($1\%$, 2 in 204)(p=0.016). Third, sidedness of resection affects to operative mortality. Right pneumonectomy ($6.8\%$, 10 in 148) showed higher operative mortality than that of left pneumonectomy ($0.8\%$, 2 in 238)(p=0.002). The group over 60 years showed higher incidence of respiratory morbidity ($11.0\%$, 20 in 182) than that of the group under 60 years ($3.4\%$, 7 in 204)(p=0.005). Right pneumonectomy also showed significantly higher incidence ($11.5\%$, 17 in 148) than that of left pneumonectomy ($4.2\%$, 10 in 238)(p=0.008). Conclusion: Age and sidedness of pneumonectomy were the risk factors of operative mortality and respiratory complications, Therefore, careful selection of patients and more attention perioperatively were demanded in right pneumonectomy. However, because the operative mortality is acceptable, pneumonectomy could be done safely if the pneumonectomy is necessary for curative resection of lung cancer.

Analysis on the Decrease of Planning Target Volume in the Case of Lung Radiation Surgery with the Application of Respiratory Gated Radiotherpy Method (폐암 환자의 방사선수술 시 호흡연동 방사선치료 방법의 적용을 통한 계획용표적체적 감소 효과 분석)

  • Song, Ju-Young;Nah, Byung-Sik;Chung, Woong-Ki;Ahn, Sung-Ja;Nam, Taek-Keun;Yoon, Mee-Sun;Jung, Jae-Uk
    • Progress in Medical Physics
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    • v.19 no.4
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    • pp.263-268
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    • 2008
  • The application of a respiratory gated radiotherpy method to the lung radiation surgery was evaluated compared with the conventional method in which the whole tumor motion range is considered in the delineation of PTV (Planning target volume). The four dimensional CT simulation images were acquired for the five NSCLC (Non-small cell lung cancer) patients for radiation surgery. The respiratory gated plan was prepared with the 50% phase CT images and the conventional method was planned based on the ITV (Internal target volume) which include all the target volumes created in each phase CT images within a whole respiratory period. The DVH (Dose volume histogram) of OAR (Organ at risk) which calculated in each method was compared for the evaluation of the plan properness. The relative decrease of OARs' DVH were verified in the application of respiratory gated method. The average decrease rate were $16.88{\pm}9.97%$ in the bronchus, $34.13{\pm}19.15%$ in the spinal cord, $28.42{\pm}18.49%$ in the chest wall and $32.48{\pm}16.66%$ in the lung. Based on these results, we can verified the applicability and the effectiveness of the respiratory gated method in the lung radiation surgery.

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Comparison of Outcomes after Curative Resection of Primary Lung Cancer between 50 Year or Younger and 70 Year or Older Patients (50세 이하와 70세 이상 원발성 폐암 환자에서의 근치적 수술 후 성적 비교)

  • Lee, Jae-Ik;Kim, Keun-Woo;Park, Kook-Yang;Park, Chul-Hyun;Jeon, Yang-Bin;Choi, Chang-Hyu
    • Journal of Chest Surgery
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    • v.42 no.2
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    • pp.206-213
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    • 2009
  • Background: Previous series have suggested that younger patients with primary lung cancer exhibit a more aggressive disease course with a worse prognosis, as compared to older patients, although this issue is still debatable. Material and Method: We reviewed the medical records of 79 patients (32 patients 50 years and younger (Group I) and 47 patients 70 years and older (Group II)) who underwent curative resection for primary lung cancer between July 2000 and June 2008. Result: The median age of the patients was 46.5 years in Group I and this was 73 years in Group II. The older patients were more likely to have major comorbidities (44% versus 77%, respectively; p=0.003). Histological examinations identified that the minor histological types (excluding non-small cell lung cancer (NSCLC)) were predominantly found in the Group I patients (16% versus 2%, respectively; p=0.037). For the TNM staging of the NSCLC, with excluding the minor histologic types, a higher proportion of patients had stage III disease in Group I (33% versus 13%, respectively; p=0.038). There was no significant difference in major morbidity (16% versus 30%, respectively; p=0.148) and operative mortality (0% versus 4.3%; p=0.512) between the groups. The mean follow-up interval was 33 months (range: $1{\sim}98$ months) for patients in both groups. For the patients with NSCLC, the five-year overall survival rate was 52.3% for Group I and 53.7% for Group II (p=0.955). The rate of freedom from recurrence at five years was significantly lower for the Group I patients than for the Group II patients (39.4% versus 70.4%, respectively; p=0.027), and only being a member of Group I impacted recurrence, based on the Cox proportional hazard analysis (p=0.034). Of the patients who had recurrence, four patients in Group I underwent aggressive surgical treatment. All of these patients exhibited long-term survival (range: $46{\sim}87$ months). Conclusion: In our study, the early outcome and long-term survival were similar for the younger and older patients after curative resection of primary lung cancer. However, we think that younger patients require meticulous follow-up as they had a tendency to proceed to surgery with advanced stage disease, a higher recurrence rate than did the older patients and the survival rates were improved, even for the recurred cases, with early aggressive treatment.